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Porcine circovirus type II gene engineering subunit vaccine, and preparation method and application thereof

A porcine circovirus and vaccine technology, applied in antiviral agents, viral antigen components, chemical instruments and methods, etc., can solve problems such as difficulties and poor virus proliferation ability, and achieve low cost, strong immunogenicity and high antigenicity. Effect

Active Publication Date: 2013-07-17
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because PCV2 does not produce cytopathic changes when cultured in vitro, and the ability of virus proliferation is poor, it is very difficult to prepare the virus vaccine by traditional methods.

Method used

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  • Porcine circovirus type II gene engineering subunit vaccine, and preparation method and application thereof
  • Porcine circovirus type II gene engineering subunit vaccine, and preparation method and application thereof
  • Porcine circovirus type II gene engineering subunit vaccine, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0053] One, the construction of the cloning vector of PCV2cap protein gene

[0054] 1 Materials and methods

[0055] 1.1 Extraction of disease material DNA

[0056] Take 0.5 g of the inguinal lymph node sample of dead pigs infected with porcine circovirus, add 4.5 ml of sterile saline, and place it in a glass homogenizer to make a homogenate. Take 472 μl lymph node homogenate, add 25 μl 10% SDS and 2.5 μl proteinase K (25 mg / ml), put it in a water bath at 50°C for 2 hours, add 500 μl water-saturated phenol, vortex for 30 seconds, and centrifuge at 12000 rpm for 10 minutes. Transfer the supernatant to another centrifuge tube, add 500 μl of phenol:chloroform:isoamyl alcohol (volume ratio 25:24:1) and mix, then centrifuge at 12,000 rpm for 10 minutes. Take the supernatant, add 3mol / L sodium acetate (pH value 5.3) according to 1 / 10 volume, add 2.5 times the volume of absolute ethanol, and put it in a -20°C refrigerator for 30 minutes. Centrifuge at 15,000 rpm for 10 minutes, di...

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Abstract

The invention discloses a porcine circovirus type II gene engineering subunit vaccine, and a preparation method and application thereof. The porcine circovirus type II vaccine comprises a soluble fusion protein composed of a protein obtained by expressing a porcine circovirus type II capsid protein gene by Escherichia coli; and the porcine circovirus type II capsid protein gene is subjected to positioning signal zone shear and site-directed mutagenesis, and the site-directed mutagenesis comprises mutation of codon AGA or AGG to CGC. A porcine circovirus type II vaccine prepared by the technical scheme provided by the invention has high antigen purity, good safety, strong immunogenicity, and no pathogenicity to animals like pig. Furthermore, the vaccine antigen in the invention is expressed by Escherichia coli, so a preparation process is relatively simple and low-cost.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to a preparation method and application of a porcine circovirus type II genetically engineered subunit vaccine. Background technique [0002] Postweaning multisystemic wasting syndrome (PMWS) was reported by John Harding of Canada in 1991, and porcine circovirus type II (Porcine circovirus 2, PCV2) was confirmed to be the main pathogen of the syndrome. [0003] Existing studies have shown that porcine circovirus disease (PCVD) is a clinical syndrome caused by PCV2 infection, and its main symptoms are progressive emaciation, shortness of breath, anemia, diarrhea, jaundice, intermittent Pneumonia, lymphadenitis and nephritis. Porcine circovirus type II mainly affects weaned piglets aged 5-12 weeks, and is closely related to PMWS, porcine dermatitis and nephritis syndrome (porcine dermatitis and nephritis syndrome, PDNS), porcine respiratory disease complex, PRDC), type A2...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/63C12N1/21A61K39/12A61P31/20
Inventor 荣俊杜元钊范根成韩建文蔡联燊
Owner YEBIO BIOENG OF QINGDAO
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